Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma and bronchioalveolar carcinoma pleural effusion tissue labelling RPL7 with ab72550 at 1/1000 (0.2µg/ml) and 1/200 (1µg/ml). Detection: DAB.
Western blot - Anti-RPL7 antibody (ab72550)This image is courtesy of an Abreview submitted by Loren Gibson
All lanes : Anti-RPL7 antibody (ab72550)
Lane 1 : U2OS whole cell lysate treated with control siRNA Lane 2 : U2OS whole cell lysate treated with RPL7 siRNA
Lysates/proteins at 10 µg per lane.
Secondary HRP-conjugated donkey anti-rabbit IgG polyclonal at 1/1 dilution Developed using the ECL technique
All lanes : Anti-RPL7 antibody (ab72550) at 0.04 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : HeLa whole cell lysate at 5 µg Lane 4 : 293T whole cell lysate at 50 µg Lane 5 : NIH3T3 whole cell lysate at 50 µg
Predicted band size : 29 kDa Observed band size : 29 kDa
Immunoprecipitation - RPL7 antibody (ab72550)
Detection of Human RPL7 by Immunoprecipitation from Whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded), using ab72550 at 3 µg/mg lysate for IP and at 1 µg/ml for subsequent Western blot detection.
IHC image of ab72550 staining in human normal cervix formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab72550, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ICC/IF image of ab72550 stained HeLa cells. The cells were 4% paraformaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab72550, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.